CORE--HEMATOPOIESIS LABORATORY

曼哈顿综合体提出的几个项目的中心目标是 镰状中心是调节γ和/或β球蛋白的合成 从患者的造血细胞的程序，以改善 镰状细胞病的症状温伯格博士的具体目标 造血核心实验室主要有：1）提供组织培养方法 并支持各组成项目。组织培养方法与技术 核心实验室建立并广泛使用的方法包括： 用于评价造血祖细胞的甲基纤维素培养物 (HPC)，B）用于逆转录病毒介导的基因的短期离体培养 干-祖细胞（S/PC）的转导和扩增，c）长期 逆转录病毒介导的S/PC基因转导的培养物， S/PC和长期培养物起始细胞（LTC-IC）的测定，以及2） 在来自以下的转导细胞中进行珠蛋白链合成研究： 培养物、移植有转导细胞的小鼠和转基因动物 用聚丙烯酰胺凝胶电泳和荧光照相法。 项目、实验和核心的作用如下： 1)项目PI：G.阿特韦实验：HPC的转换和扩展， 分析来自转导细胞的集落， 转导的细胞和集落。核心的作用：短期流动性 转导培养，两相液体培养，甲基纤维素培养 以及凝胶电泳和荧光照相。 2)项目PI：A.银行实验：HPC的转换和扩展。 来源于长期培养物的集落中的转导分析- 启动细胞（LTC-IC）和HPC，分析细胞的珠蛋白合成 和殖民地。核心的作用：长期骨髓培养+/-基质，+/- 造血生长因子（HGF），短期转导培养物+/- 基质、凝胶电泳和荧光照相。 3)项目PI：F. Constantini：实验：珠蛋白链分析 在转基因小鼠中的合成。核心作用：网织红细胞 孵育、凝胶电泳、荧光照相。 4)项目PI：J.Bieker，实验：用EKLF样锌培养HPC 手指蛋白，分析珠蛋白合成的细胞和集落，从这些 cultures.核心的作用：甲基纤维素和两相液体培养物，凝胶 电泳荧光法 5)项目PI：S.皮奥梅利实验：HPC培养物+/-精氨酸 丁酸盐，体内精氨酸丁酸盐前、期间和后，检查 一个接一个地合成珠蛋白。核心作用：甲基纤维素 以及凝胶电泳和荧光照相。

A central goal of several projects proposed by the Manhattan Comprehensive Sickle Center is to modulate the gamma- and/or beta globin synthesis program of hematopoietic cells from patients in order to ameliorate the symptoms of sickle cell disease. The specific aims of Dr. Weinberg's Hematopoiesis Core Laboratory are: 1) To provide tissue culture methods and support to component projects. Tissue culture methods and techniques which are established and used extensively in the core lab include: a) methylcellulose cultures for evaluation of hematopoietic progenitor cells (HPC), b) short term ex vivo cultures for retroviral mediated gene transduction and expansion of stem-progenitor cells (S/PC), c) long term cultures for retroviral mediated gene transduction of S/PC, expansion of S/PC, and assay of long term cultures initiating cells (LTC-IC), and 2) To perform globin chain synthesis studies in transduced cells derived from cultures, mice transplanted with transduced cells, and transgenic animals using polyacrylamide gel electrophoresis and fluorography. Projects, experiments, and core's role are as follows: 1) Project PI: G. Atweh. Experiments: Transduction and expansion of HPC, analysis of colonies derived from transduced cells, globin synthesis by transduced cells and colonies. Role of core: short term liquid transduction cultures, two phase liquid cultures, methylcellulose cultures and gel electrophoresis and fluorography. 2) Project PI: A. Bank. Experiments: Transduction and expansion of HPC. Analysis of transduction in colonies derived from long term culture- initiating cells (LTC-IC) and HPC, analysis of globin synthesis by cells and colonies. Role of core: Long term marrow cultures +/-stroma, +/- hematopoietic growth factors (HGFs), short term transduction cultures +/- stroma, gel electrophoresis and fluorography. 3) Project PI: F. Constantini:Experiment: Analysis of globin chain synthesis in transgenic mice.colonies. Role of core: Reticulocyte incubations, gel electrophoresis, fluorography. 4) Project PI: J.Bieker,Experiments: Culture HPC with EKLF-like zinc finger proteins, analyze globin synthesis in cells and colonies from these cultures. Role of Core: Methylcellulose and two phase liquid cultures, gel electrophoresis and fluorography 5) Project PI: S. Piomelli. Experiments: HPC cultures +/-arginine butyrate, pre-, during, and post- in vivo arginine butyrate, examine globin synthesis by colonies by colonies. Role of core: Methylcellulose and gel electrophoresis and fluorography.